changes in protein profiles in bortezomib applied multiple myeloma ...

3.2.5. Cell Proliferation Assay
Anti-proliferative effects of Bortezomib on U-266 MM cells were determined by
XTT cell proliferation assay. The principle of this assay is based on the ability of
metabolic active cells to reduce the tetrazolium salt XTT to orange colored compounds
of formazan by their mitochondrial enzyme. These formazan molecules formed is water
soluble and can be measurable at the wavelength of 492 nm with a spectrophotometer.
Briefly, the test procedure includes cultivation of cells in a 96-well plate, adding
the XTT reagent (The procedure for preparation of XTT Reaction Solution that was
used in XTT Assay was given in Appendix B) and incubation for 72 hours. During
incubation an orange color is formed. The greater the number of the active cells in the
well, the greater the activity of mitochondria enzymes and the higher the concentration
of the formazan compounds (dye) formed, which can then be measured and quantitated
with a spectrophotometer.
2x10 4 U-266 cells (100 µl) were seeded into 96-well plates containing 200 µl
growth medium in the absence or presence of increasing concentrations of Bortezomib
(Figure 3.1)
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96 Well Plate A
Figure 3.1. The Dosage of Bortezomib (nM) Applied on MM U-266 Cells
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